丁香实验_LOGO
登录
提问
我要登录
|免费注册
点赞
收藏
wx-share
分享

Use of Antisense-Generating Plasmids to Probe the Function of Signal Transduction Proteins in Primary Neurons

互联网

532
Investigation of the function of intracellularly located protein components of receptor-effector transduction pathways has been hampered not only by their inaccessibility, but also by the lack of specific tools. The problem is particularly acute when the effect of interest must be measured at the single cell level, for example, receptor modulation of ion channels. Assignment of different G protein subunits to roles in ion channel modulation by receptors progressed greatly with the availability of antibodies raised against peptide sequences corresponding to the carboxyl-terminus of Gα-subunits. This is the part of the a-subunit molecule thought to interact with effector, so these antibodies were predicted to indirectly occlude receptor-effector interaction. Indeed, we found that injection of an antibody against Gα0 , into sympathetic neurons inhibited α2 -adrenoceptor effects on Ca2+ current, and antibody against Gαq/11 inhibited muscarinic and bradykinin receptor effects on the M-type K+ current (1 ,2 ). However, this approach is seriously limited by the close sequence homologies between members of G protein a-subunit subfamilies (3 ), which results in the antibodies being unable to discriminate between members in many cases.
ad image
提问
扫一扫
丁香实验小程序二维码
实验小助手
丁香实验公众号二维码
扫码领资料
反馈
TOP
打开小程序